Medicine
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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty
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Item Detection of human strongyloidiasis among patients with a high risk of complications attending selected tertiary care hospitals in Colombo, Sri Lanka using molecular and serological diagnostic tools(BioMed Central, 2024-10) Weerasekera, C.J.; Gunathilaka, N.; Menike, C.; Anpahalan, P.; Perera, N.; De Silva, N.R.; Wickremasinghe, R.BACKGROUND Strongyloidiasis a neglected tropical disease is known to cause severe disease among immunosuppressed and has not been studied extensively in Sri Lanka. Parasitological diagnostic approaches based on faecal microscopy and culture often fail to detect low-intensity infections. This study investigates the presence of strongyloidiasis among selected immunocompromised individuals using parasitological, molecular and serological techniques.METHODS Adult patients with immunocompromising conditions admitted to three tertiary care hospitals in Sri Lanka were recruited. A faecal sample and 2 ml of venous blood were collected. The faecal samples were subjected to direct faecal smear and cultures (agar plate, charcoal and Harada-Mori) and polymerase chain reaction (PCR) using species specific primers designed for Strongyloides stercoralis. The presence of Strongyloides IgG antibodies was tested in the collected serum samples using DRG Strongyloides IgG enzyme-linked immunosorbent assay (ELISA) kits. The PCR products of the positive samples were sequenced using Sanger sequencing method.RESULTS A total of 260 patients were recruited to this study, out of which 160 provided faecal samples and 122 provided blood samples. Out of the 160 faecal samples, none were positive for strongyloidiasis by direct smear, charcoal and Harada-Mori cultures. Only one sample (0.6%) was positive by agar plate culture. Out of the 123 samples subjected to PCR, 14 (11.4%), including the culture positive patient, were positive for S. stercoralis. Sequencing results of the PCR products indicated 100% similarity to S. stercoralis. Out of the 122 serum samples subjected to ELISA, 20 (16.4%), including the culture positive patient, were positive for Strongyloides IgG antibodies. However, sociodemographic, exposure factors, clinical features were not significantly associated with the presence of strongyloidiasis infection.CONCLUSIONS Strongyloidiasis is present among the immunocompromised population in Sri Lanka, even in the absence of a significant relationship with associated factors. It is advisable to screen such patients with highly sensitive tests such as PCR for early diagnosis and treatment.Item Abundance and taxonomic characterization of chigger mites (Acari: Trombiculidae and Walchiidae) associated with rodents in selected scrub typhus-prone areas in Southern and Western provinces of Sri Lanka(University of Kelaniya, 2024) Liyanage, A.; Gunathilaka, N.; Premarathne, R.; Chandrasena, N.; Jacinavicius, F.D.C.; Silva, R.B.Larval trombiculid mites (chiggers) are the vectors and reservoirs of the potentially lethal infectious disease, scrub typhus (ST) caused by Orientia tsutsugamushi. Small rodents are natural hosts of parasitic larval stage of the chigger mites. This study focused on determining the abundance of chigger mites associated with rodents in Sri Lanka and the taxonomic characterization of field-caught chiggers. Field sampling was conducted in the districts of Galle, and Hambantota of the Southern Province, and Gampaha of the Western Province, in 2019 and 2020. Sampling sites were selected according to the patient distribution. Rodents were captured using baited traps (7.62 cm x 7.62 cm x 25.4 cm) set up just before sunset at peri-domestic or work premises of ST fever patients. A total of 422 traps were placed at identified possible exposure locations in Galle (n=122), Hambantota (n=120) and Gampaha (n=178). A total of 58 small mammals were captured [Galle (n=19), Hambantota (n=7), and Gampaha (n=32)] under three small rodent species, namely; Rattus rattus (Black rat), Rattus norvegicus (Brown rat), Tatera indica (Indian gerbil), Gollunda ellioti (Indian bush rat) and Suncus murinus (Asian house shrew). The trapped rodents were anaesthetized with ketamine (75 mg/kg) /xylazine (10 mg/kg) and examined for larval mites. Mites detected were removed carefully with a brush, collected, and washed individually with 10% PBS and slidemounted in Hoyer’s medium. Chiggers were speciated morphologically by visual inspection and morphometry using a camera-mounted light microscope (x100). A total of 394 life stages of mites were collected. Three different genera were identified, including Leptotrombidium, Schoengastiella, and Microtrombicula. Leptotrombidium imphalum (72.59%; n=286) was the predominant species, followed by Schoengastiella punctata (8.12%; n=32). Some specimens were identifiable only up to genus level, Leptotrombidium sp. (3.55%; n=14) and Microtrombicula sp. (4.82%; n= 19). Some (7.11%; n=28) were not trombiculid mites, while 3.81%; n=15 was damaged beyond identification. Leptotrombidium imphalum was detected for the first time parasitizing the murids - Rattus novergicus and Tatera indica in the district of Galle, a new locality. In addition, S. punctata was recorded in a new locality in the Gampaha district, Western province with a new host association, Golunda ellioti. This study emphasizes the need for further entomological surveys in ST disease-endemic areas. Developing a morphological identification key for chigger mites in Sri Lanka is a top priority to facilitate field surveys.Item Parasitological screening of vector mosquitoes and molecular biological identification of larval filarial parasites among the wildcaught Mansonia mosquito species at selected areas in the district of Gampaha, Sri Lanka, a re-emerging focus of Brugian filariasis(University of Kelaniya, 2024) Gunathilaka, N.; Wimalasiri, U.; Chandrasena, N.; Dalpadadu, R.Brugian filariasis, a disease caused by the Protozoan parasiteBrugia malayi has re-emerged in Sri Lanka after nearly four decades of quiescence. The Brugia malayi that prevailed in Sri Lanka in the past was the nocturnal periodic human strain transmitted by mosquitoes of the genus Mansonia. The objective of the present study was the precise identification of vector mosquitoes and parasites of the current onset of the disease. Entomological surveys were performed during September/October 2021 in Ragama Medical Officer of Health area using cattle-baited net traps. Mansonia sp. mosquitoes were dissected to detect the presence of larvae of the parasite. The lysate of dissected mosquitoes positive for larvae was used for the extraction of genomic DNA of the parasite, which was subjected to Polymerase Chain Reactions (PCR) aimed at molecular speciation using pan-filarial primers specific for the internal transcribed spacer region two (ITS2) of the ribosomal DNA. A total of 1060 mosquitoes were tested, and that included seven mosquito species belonged to four genera. Culex gelidus (n=602; 56.8%) was detected as the predominant mosquito species followed by Armigeres subalbatus (n=420; 39.6%) Cx. tritaeniorynchus (n=2; 0.2%) and Anopheles nigerrimus (n=4; 0.4%). Mansonia spp. accounted for 2,7% of the total mosquito sample and among them, the presence of Mansonia annulifera was 1,2% of the total (n=20), Ma. uniformis was 0.9% (n= 10) and Ma. Indiana was 0.2% (n= 2). About 18.7% (n=6) of Mansonia mosquito collection was positive for filarial larvae. Among them, 15.6% (n=5) was Mansonia annulifera while (3.1%; n=1) was Ma. uniformis. The PCR products of all tested samples corresponded to the band size of 625 bp, specific to B. malayi confirming the identity of the parasite. Mansonia annulifera and Ma. uniformis were confirmed as vectors of the re-emerged B. malayi (nocturnally sub-periodic) in Gampaha district. The role of other mosquito vector species would require investigation by vector incrimination and xenomonitoring-based approaches.Item Forecasting dengue incidence based on entomological indices, population density, and meteorological and environmental variables in the Gampaha District of Sri Lanka(Elsevier Ltd, 2024) Dalpadado, R.; Amarasinghe, D.; Gunathilaka, N.; Wijayanayake, A.N.No abstract availableItem The presence of strongyloidiasis and associated risk factors in patients undergoing treatment at the National Cancer Institute, Maharagama, Sri Lanka(Sri Lanka Medical Association, 2023) Weerasekera, C.J.; Menike, C.W.; Wimalasiri, U.; Wijerathna, T.; Jayathilake, D.C.C.; Somawardane, U.A.B.P.; Saravanamuttu, U.; Yoganathan, N.; Perera, N.; Gunathilaka, N.; de Silva, N.R.; Wickremasinghe, D.R.INTRODUCTION: Strongyloides stercoralis can cause severe disease in the immunocompromised. Without a proper gold-standard diagnostic technique, strongyloidiasis is scarcely studied both globally and locally. OBJECTIVES: We aimed to estimate the prevalence of strongyloidiasis among immunocompromised adult patients and to identify risk factors. METHODS: This study was carried out between February to October 2022. A faecal sample and 2 ml of venous blood were collected from consented patients. Direct faecal smear, agar plate, Harada-Mori and Charcoal cultures were performed on the faecal samples. Qualitative Polymerase Chain Reaction (PCR) was performed on selected faecal samples using S. stercoralis targeting ITS1 region. Strongyloides IgG ELISA was carried out on the serum samples using DRG Strongyloides IgG ELISA kit. RESULTS: Collectively, 144 patients (males = 68, females = 76) provided blood/faecal sample or both. Relevant to strongyloidiasis-associated symptoms, some patients had diarrhoea (n=12) and eosinophilia (n=11). Some of them (n=74) had occupational or recreational exposure to soil as potential risk factors. Overall, 24 patients were positive for strongyloidiasis from one or more diagnostic method (5 PCR and 19 ELISA). There were zero culture or direct smear positives. There was no significant association between disease positivity with either of the clinical features or risk factors. CONCLUSION: The prevalence of strongyloidiasis in patients with malignancies was 16.66%. Strongyloidiasis is existent in the immunocompromised in Sri Lanka even in the absence of suggestive clinical features or regular exposure to risk factors. Screening immunocompromised patients with sensitive techniques such as PCR for timely diagnosis and treatment is recommended.Item Ocular trematodiasis in children, Sri Lanka(National Center for Infectious Diseases, 2023) Mallawarachchi, C.H.; Dissanayake, M.M.; Hendavitharana, S.R.; Senanayake, S.; Gunathilaka, N.; Chandrasena, T.G.A.N.; Yahathugoda, T.C.; Wickramasinghe, S.; de Silva, N.R.Using histopathology and phylogenetic analysis of the internal transcribed spacer 2 gene, we found >2 distinct trematode species that caused ocular trematode infections in children in Sri Lanka. Collaborations between clinicians and parasitologists and community awareness of water-related contamination hazards will promote diagnosis, control, and prevention of ocular trematode infections.Item Genetic diversity of Leishmania donovani isolates from cutaneous lesions of military personnel in the Mullaitivu and Kilinochchi districts of the Northern Province, Sri Lanka(Wolters Kluwer, 2022) Wijerathna, T.; Gunathilaka, N.; Semege, S.; Pathirana, N.; Rodrigo, W.; Fernando, D.Objective: To compare the DNA sequences of Leishmania (L.) donovani isolated from individuals in two districts of the Northern Province with other parts of Sri Lanka and neighboring countries. Methods: Samples were collected from military personnel at the Army Hospital, Narahenpita, Sri Lanka from November 2018 to March 2020. A portion of the samples was fixed, stained with Giemsa and observed under the light microscope. The genomic The DNA was extracted from the remaining portion of the samples using DNEasy blood tissue kit (Qiagen, Germany) and amplified using Leishmania genus-specific primers for molecular diagnosis initially. DNA was amplified using L. donovani species-specific primers by PCR and the amplified product was sequenced for comparison of nucleotide sequences. Results: Out of 76 suspected patients, at least one biological sample of 45 (59.2%) was positive for L. amastigotes upon microscopy. Overall, 33 (43.4%) were positive in Leishmania genus-specific PCR, but only 23 (30.3%) were positive in L. donovani specific PCR. The dendrogram indicates that the current sequences clustered together with those from Nepal and Gampaha districts (Western Province), Sri Lanka, while the Indian and Eastern African sequences clustered separately. Conclusions: The genetic diversity was low among the isolates, indicating a single and possibly a local point of origin. However, the similarity of Sri Lankan and Nepal strains indicate a possibility of a shared point of origin, which needs more extensive evidence to confirm.Item Time series analysis of leishmaniasis incidence in Sri Lanka: evidence for humidity-associated fluctuations(Springer Verlag, 2023) Wijerathna, T.; Gunathilaka, N.Leishmaniasis is a vector-borne disease of which the transmission is highly influenced by climatic factors, whereas the nature and magnitude differ between geographical regions. The effects of climatic variables on leishmaniasis in Sri Lanka are poorly investigated. The present study focused on time-series analysis of leishmaniasis cases reported from Sri Lanka with selected climatic variables. Variance stabilized time series of leishmaniasis patients of entire Sri Lanka and major districts from 2014 to 2018 was fitted to autoregressive integrated moving average (ARIMA) models. All the possible models were generated by assigning different values for autoregression and moving average terms using a function written in R statistical program. The top ten models with the lowest Akaike information criterion (AIC) values were selected by writing another function. These models were further evaluated using RMSE and MAPE error parameters to select the optimal model for each area. Cross-autocorrelation analyses were performed to assess the associations between climate and the leishmaniasis incidence. Most associated lags of each variable were integrated into the optimal models to determine the true effects imposed. The optimal models varied depending on the area. SARIMA (0,1,1) (1,0,0)12 was optimal for the country level. All the forecasts were within the 95% confidence intervals. Humidity was the most notable factor associated with leishmaniasis, which could be attributed to the positive impacts on sand fly activity. Rainfall showed a negative impact probably as a result of flooding of sand fly larval habitats. The ARIMA-based models performed well for the prediction of leishmaniasis in the short term.Item Genetic variation of sand flies (Diptera: Psychodidae) in Gampaha and Kurunegala districts of Sri Lanka: Complementing the morphological identification(Wolters Kluwer Medknow Publications, 2022) Wijerathna, T.; Gunathilaka, N.; Rodrigo, W,OBJECTIVE: To identity the variation of sand flies in the Gampaha and Kurunegala districts of Sri Lanka and to assess DNA barcoding as a complementing method for morphological identification. METHODS: A total of 38 441 sand flies were collected from selected localities in Gampaha and Kurunegala districts using standard entomological techniques from May 2017 to December 2018. Specimens were identified using morphological features and compared with mitochondrial cytochrome C oxidase subunit I gene- based DNA barcoding as an alternative tool. RESULTS: Morphological and molecular identification confirmed the presence of four species under two genera (Phlebotomus and Sergentomyia). Phlebotomus argentipes was the predominant species, followed by Sergentomyia (S.) punjabensis, S. babu insularis, and an unidentified Sergentomyia sp. Phlebotomus argentipes showed a clear genetic differentiation from other species. S. babu insularis and S. punjabensis showed a higher genetic affinity to each other than the unidentified species. The unidentified Sergentomyia species is morphologically similar to S. zeylanica, but differs only in clavate gonostyle. CONCLUSIONS: DNA barcoding is an effective technique for the identification of sand flies. Further studies using molecular techniques will improve the knowledge of the cryptic diversity of Sri Lankan sand fly fauna. Establishing a reliable and standardized identification system for sand fly species in Sri Lanka is recommended.Item Public knowledge, practices and perceptions on typhus fevers in Southern Sri Lanka(Wolters Kluwer Medknow Publications, 2022) Liyanage, A.; Chandrasena, N.; Gunathilaka, N.; Sanjeewa, R.; Premaratna, R.Objective: To assess public knowledge, practices and perceptions on typhus fevers in Sri Lanka. Methods: A descriptive study was done in four selected typhus- prone areas in Southern Sri Lanka. A mixed-method was employed using face-to-face interviews and questionnaire-based surveys among confirmed cases of typhus and at-risk populations, respectively. Frequencies, percentages, and means were used to characterize socio-demography and evaluate disease awareness. Results: The lay terms for typhus fevers reported in the studied region were “peacock fever”, “tick fever” and “bird fever”. A total of 499 subjects participated [mean±SD, (45±16) years] in the questionnaire-based survey, and 13.6% (n=68) reported past experience of typhus fever, 1.2% (n=6) identified the disease as “typhus” while 58.7% (n=293) and 11.8% (n=59) knew it as ‘peacock fever’ and ‘tick fever’, respectively. The etiological agent was unknown to 95.2% (n=475), but 53.5% ((n=267) were aware that it was vector-borne. Fever (57.3%, n=286), eschar (35.7%, n=178), headache (22.0%, n=267) and myalgia (19.2%, n=96) were identified as key symptoms. Past disease experience was significantly associated with higher awareness of the main disease symptoms (fever: χ2=15.713, P<0.001; headache: χ2=19.447, P<0.001; lymphadenopathy: Fisher’s exact test, P=0.023; eschar: χ2=12.049, P<0.001). None knew of any disease prevention methods. Participants with a past history of typhus fever had sought treatment at state hospitals (55.9%, 38/68) and private sector hospitals (5.9%, 4/68). Conclusions: Public awareness on preventive practices for typhus fevers was rare among the participants though vector-borne aspect was known to many. Clinical disease awareness was deficient among those without past experience of typhus fever. Community sensitization on vector avoidance strategies is highly recommended.